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Identification of novel catechin receptor that mediates blood vessel protective effect and vasodilatation, and it's application for functional food

介导血管保护作用和血管舒张作用的新型儿茶素受体的鉴定及其在功能食品中的应用

基本信息

批准号：
18580113
负责人：
NAKAI Yuji
金额：
$ 2.5万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580113/
关键词：
food physiology proteomics receptor polyphenol

项目摘要

From preliminary investigation, we found a vasodilatation effect of (+)-catechin on the endothrial cells. The effect was observed within 30 minutes after (+)-catechin treatment in vivo, but not (-)-epigallocatechin gallate, we hypothesized that the existence of novel (+)-catechin specific receptor and that the effect was mediated through the receptor signaling thereby produced NO. In order to identify putative (+)-catechin receptor, we used a differential proteomics approach in the brain of (+)-catechin injected mice. Male, 8-week-old ICR mice were injected 100 mg/ml (+)-catechin intravenously. After 3h injection, the brain was excised and homogenized to extract proteins. The brain extracts were subjected to 2-D PAGE, and then the 2-D gels were silver stained. The spots with a pI of 〜8.1 and a molecular mass of 〜60 kDa and a pI of 〜8.3 and a molecular mass of 〜17 kDa were markedly increased in (+)-catechin injected mouse brain extract compared with vehicle injected control. While the spot with a pI of 〜5.7 and a molecular mass of 22 kDa was decreased. These changes in spots are suspected to be brought about by the post-translational modification such as protein phosphorylation or protein degradation of the putative receptor itself or its downstream signaling molecules. Next, we will try to isolate these protein spots and analyze the amino acid sequences using peptide mass fingerprinting.

初步研究发现，（+）-儿茶素对血管内皮细胞有舒张作用。在体内（+）-儿茶素处理后30分钟内观察到该作用，但（-）-表没食子儿茶素没食子酸酯没有观察到，我们假设存在新的（+）-儿茶素特异性受体，并且该作用通过受体信号传导介导，从而产生NO。为了鉴定推定的（+）-儿茶素受体，我们在注射（+）-儿茶素的小鼠的脑中使用差异蛋白质组学方法。将8周龄的雄性ICR小鼠静脉内注射100 mg/ml（+）-儿茶素。注射3 h后，切除脑并匀浆以提取蛋白质。脑组织提取液进行双向电泳，然后进行银染。与注射载体的对照相比，在注射（+）-儿茶素的小鼠脑提取物中，pI为8.1和分子量为1060 kDa以及pI为8.3和分子量为1017 kDa的斑点显著增加。而pI为5.7，分子量为22 kDa的斑点减少。这些斑点的变化被怀疑是由翻译后修饰引起的，例如推定受体本身或其下游信号分子的蛋白质磷酸化或蛋白质降解。接下来，我们将尝试分离这些蛋白质点，并使用肽质量指纹法分析氨基酸序列。